GSM7178946: male, 28 degrees, biorep2; Drosophila melanogaster; RNA-Seq
RNA was wxtracted with TRIzol following manufacturer protocol. 1.mRNA enrichment and purification. Oligo dT Selection (mRNA enrichment): Use oligo dT beads toenrich mRNA with poly A tail.   2.RNA fragment and reverse transcription.RNA fragmentation and first-strand cDNAs were generated using random N6-primed reverse transcription, followed by the second-strand cDNA synthesis.   3.End repair, add A and adaptor ligation.The synthesized cDNA was subjected to end-repair and then was 3' adenylated. Adaptors were ligated to the ends of these 3' adenylated cDNAfragments.   4.PCR amplification. Many rounds of PCR amplification were performed to enrich the purified cDNA template using PCR primer.   5.Cyclization. Denature the PCR product by heat and the DNA iscyclized by splint oligo and DNA ligase.   6.DNA nanoball synthesis.